Flow cytometric measurement of reactive oxygen species production in red blood cells from fish after exposure to heavy metals

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Article Title:	Flow cytometric measurement of reactive oxygen species production in red blood cells from fish after exposure to heavy metals
Authors:	Aurelia Covaci, Ciprian-Valentin Mihali, Violeta Turcuş, Daniela Bratosin
Affiliation:	Faculty of Natural Sciences, Engineering and Informatics, ″V. Goldis″ Western University of Arad, Romania Institute of Life Sciences, ″Vasile Goldis″ Western University of Arad, Romania National Institute for Biological Science Research & Development (INCDSB), Bucharest, Romania
Abstract:	Organisms react to environmental pollutants by disturbance of living processes at subcellular levels resulting in cells death. To understand the mechanisms underlying the process of cell death by heavy metals action, we measured by flow cytometry the reactive oxygen species (ROS) generation by heavy metal (Al, Cd, Hg, Pb, Zn) in fish nucleated erythrocytes correlated with morphological changes by scattered light flow cytometry in the mode FSC/SSC. Optical and scanning electron microscopy were accompanied the research. The results we obtained show that heavy metals generates a significant amount of ROS that induce significant morphological changes characteristic for the phenomenon of cell death by apoptosis. The present results demonstrate that oxidative stress assessment and the use of fish nucleated erythrocytes “in vitro”, can be a good tool for ecotoxicological evaluation of the presence of pollutants, heavy metals in particular.
Keywords:	nucleated erythrocytes, fish heavy metals, apoptosis, reactive oxygen species, flow cytometry, ESEM, optic macroscopy, ecotoxicology
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*Correspondence:	Daniela Bratosin, National Institute for Biological Science Research & Development, Bucharest, Romania; Splaiul Independentei no. 296, district 6, Bucharest, Romania, Tel/Fax +40-(021)-2200881, email:bratosind@yahoo.com